Transcriptomics-Based Study of Immune Genes Associated with Subclinical Mastitis in Bactrian Camels.

Abstract

The significant increase in demand for camel milk has led to a rapid increase in the number of Bactrian camels. However, the widespread occurrence of mastitis significantly impacts the development of the Bactrian camel milk industry and poses a public health risk. Despite this, there is a lack of research on the transcriptional response, immune response pathways, and changes in core genes of Bactrian camels with subclinical mastitis. This study aimed to reveal the changes in immune-related response pathways and gene transcription levels in Bactrian camels with subclinical mastitis by analyzing the blood transcriptional response after the occurrence of subclinical mastitis in natural conditions. This study focused on 7-year-old Bactrian camels and collected 2 mL of blood from the camels that tested positive with a 4-peak California Mastitis Test (CMT) and those that tested negative with a 3-peak CMT. RNA sequencing (RNA-Seq) technology was used to analyze gene expression in the blood samples. Gene expression was verified using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Overall, 1722 differentially expressed genes were sequenced in the blood samples of CMT-positive and CMT-negative Bactrian camels, including 1061 upregulated and 661 downregulated genes. After conducting gene ontology functional enrichment, 453 differentially expressed genes were identified. We also discovered pathways such as immune response, the G-protein-coupled receptor signaling pathway, and internal signal transmission. Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment detected 668 differentially expressed genes annotated onto 309 metabolic pathways, with significantly enriched immune pathways including cytokine-cytokine receptor interaction, complex and coalescence cascades, natural killer cell-mediated cytotoxicity, and T helper type 17 cell differentiation, among others. Through a STRING protein interaction database and cytoscape analysis, it was found that core differentially expressed genes related to immunity included IL10, CCL5, IL1B, OSM, TNFRSF1B, IL7, and CCR3, among others. The RT-qPCR results for six randomly selected core differentially expressed genes showed that the RT-qPCR expression pattern was consistent with the RNA Seq results. The immune-related genes in Bactrian camels affected by subclinical mastitis are primarily concentrated in the immune response and the cytokine-cytokine receptor interaction pathway. Given the importance of these pathways and the connections among related genes, the immune genes within these pathways may play a crucial role in the pathogenesis of subclinical mastitis in Bactrian camels. This study provides a valuable reference for investigating the immune regulatory mechanisms of subclinical mastitis in Bactrian camels.